1. Field of the Invention
This invention relates to treating body fluid-related diseases, where pathogenic microorganisms and cells infected by the microorganisms are mainly contained in the body fluid, particularly such retrovirus diseases such as AIDS.
2. Behavior of HIV
AIDS is the acronym for Acquired Immunodeficiency Syndrome, caused by Human Immunodeficiency Virus (HIV). The behavior of HIV must be reviewed as the basis of this invention.
HIVs and HIV-infected cells in human blood, seminal fluid, vaginal mucous, etc., are spread from exchange of HIV-infected blood or lymphatic fluid or from contact with infected mucous membrane, thereby establishing a new infection of AIDS.
Glycoprotein, gp120 developed on the HIV envelope in a large quantity, adheres to CD4 protein, developed on the cell membrane of certain leucocyte cells as T-4 helper lymphocytes, macrophage and dendritic cells. These cells are known as CD4 (positive) cells. Fusion of the HIV envelope with the CD4 cell membrane is carried out by gp41 on the HIV envelope. The HIV RNA and reverse transcriptase of the HIV are brought into the CD4 cell. The HIV RNA is reverse transcribed as a complement DNA (pro virus) by the reverse transcriptase, and the provirus-DNA is combined into the host CD4 cell DNA.
The host cell then replicates the HIV RNA from the provirus-DNA as if the provirus were part of its own DNA. It further generates corresponding protein, cuts the protein into HIV components with the protease and assembles the HIV components into a new HIV. New HIVs thus bud out from the host cell membrane, mature and isolate themselves as free HIVs from the host cell. This proliferation process is repeated. HIV cannot proliferate by itself without the host cell. After proliferation is repeated, the host cell dies, reportedly because the host cell membrane has broken. However, more probably, the host cell (infected cell) dies as a result of apoptosis or the programmed death, caused by such substances as TNF or anti-Fas antibody.
When HIVs or infected cells enter human body fluid, CD4 cells are infected and the infected cells replicate free HIVs in such body fluid as blood. Within 6 to 8 weeks, the immune mechanism of human body function forms antibody to HIV, and the HIVs in the blood almost disappear. This antibody is used to verify HIV infection. However, the number of CD4 cells in blood remains nearly equal to and a little less than that of a-healthy person, around 800xcx9c1,200/mm3.
The newly infected HIV carrier then enters an asymptomatic latent period, which lasts about 5 to 10 years. During the latent, period, the number of HIVs and CD4 cells in the patient""s blood will not increase substantially. The number of infected CD4 cells remains around 0.2xcx9c1.0% of the total CD4 cells in the blood.
The activity of HIV/infected cells has been thought to be low during the latent period. But it was discovered in March 1993 that most of the HIV-infected CD4 cells remain in lymphonodes throughout the latent period and that new infection progresses during that time. Anthony S. Fauci et al., xe2x80x9cHIV infection is active and progressive in lymphoid tissue during the clinically latent stages of disease.xe2x80x9d Nature, Vol. 362, No. 6418 (Mar. 25, 1993); 355-358. The mechanism for this progressive pathology is not clear but assumed to be as follows. Healthy CD4 cells also present in a lymphonode may be infected from direct cell-to-cell contact with an infected cell. Gp120 developed on the infected cell adheres to CD4 of the healthy cell and cell fusion occurs to form a giant multinucleic cell. Healthy CD4 cells may also be infected with free HIVs generated from neighboring infected cells. As a result of cell-to-cell infection and death of the infected cell (from apoptosis or the like), the number of CD4 cells decreases at the rate of 50/mm3 every year.
When the number of CD4 cells in the blood approaches 400xcx9c300/mm3, HIV proliferation of the infected cells is activated by certain factors, and the number of HIVs and infected cells in the blood begins to increase. This stage of infection is described as AIDS Related Complex (ARC). The decrease in CD4 cells allows diseases that have been suppressed by the immune system until then an invasive opportunity. Such diseases include infections such as pneumocystis carinii, cytomegalovirus, candidosis, etc., and neoplasms such as Kaposi""s sarcoma, non-Hodgkin""s lymphoma, etc.
Almost 10% of AIDS patients also suffer from neuropathy. It has been reported but not definitively shown that macrophage, one of the CD4 cells existing in the brain, becomes infected and causes degeneration of the central nervous system, although the degeneration mechanism is not yet clear. Thus, HIVs and infected cells may exist in cerebrospinal fluid. It is also reported that dendritic cells are infected and exist in Langerhans cells under skin or tissue surfaces. Regardless, both infections may be introduced through the blood stream.
Once the number of CD4 cells in blood decreases below 200/mm3, the ARC period has evolved into AIDS. Patients may die in about one year as a result of the dominant ARC enhanced by extreme decrease of CD4 cells in blood.
It is not clear what factor causes the transition from the asymptomatic, latent period to the symptomatic (ARC,AIDS) period. Recently, it was reported(*a) that such transition occurs two months after the spontaneous mutation of 12th base in V3 region of gp120. However, it is not probable that all infected cells are activated by the same mutation. So it is natural to assume a certain substance that the inventors of the present invention call Trigger Factor initiates and stimulates the activation of infected cells and induces the asymptomatic-to-symptomatic transition. The activated infected cells may again generate the Trigger Factor and accelerate the disease. The Trigger Factor may stimulate the proliferation control region of provirus-DNA, accelerate HIV replication and finally bring the host cells to death through the cell membrane breakage or apoptosis.
It is found in vitro that TNF, namely tumor necrosis factor (and/or anti-Fas antibody, etc.) causes coagulation of DNA of the infected T-4 lymphocytes about 3 hours after administration and further subjects the infected cells to death(*b), and that HIV replication is highly stimulated during the process. TNF is not found in blood during the asymptomatic, latent period, but found during the symptomatic (ARC,AIDS) period. TNF could be one of the Trigger Factors defined by the inventors of the present invention. It is assumed(*c) that the infected cells may die following the program incorporated by human body defense system,xe2x80x94apoptosis. TNF (or anti-Fas antibody, etc.) is now the subject of research in this line.
3. Chemical Treatmentxe2x80x94Prior Art
Development of vaccine has been unsuccessful, because the identifying target of the vaccine, gp120, is subject to very rapid mutation, more specifically in V3 region, resulting in too much variety of HIV stocks (quasi-HIV species).
Various medicines are under development, inhibiting various stages of HIV infection and proliferation such as adhesion to CD4, fusion into the host cell, reverse transcription, insertion into DNA, RNA replication, protein generation, dividing the protein into components, assembling the components, budding, maturing, etc. Ten or more medicines are reported successful in vitro, but none of them has been successful in vivo. Either they have not worked or have exhibited strong toxicity as a side effect to medication. Exceptions are AZT, ddI, ddC as reverse transcription inhibitors, but they are effective only in extending the amount of time before death and also show toxicity in long-term administration, and drug-resistant HIV stocks to them are found.
4. Extracorporeal Blood Processingxe2x80x94Prior Art
Incurability of AIDS by chemical treatment has led to development of various means of extracorporeal removal, inhibition or destruction of the blood borne HIVs and infected cells. Such means originate from the technology used for conventional disease like cancer, type-B hepatitis and lymphocyte-related illnesses.
The following modes are disclosed for the extracorporeal blood processing to treat AIDS.
Processing as whole blood.
Separating into two fractions, plasma fraction and blood cell fraction and then processing the plasma fraction. This is the so-called plasmapheresis.
Separating the white blood cells and plasma fraction from the red cell fraction, after which the former fraction is processed.
Separating the leukocyte (white blood cell) fraction from the red cell fraction, after which the leukocyte fraction is processed.
Processing blood is expected to influence other body fluid systems such as lymphatic system because of intracorporeal exchange between the blood and all the other body fluids(*m).
Administering heparin before the process to induce lymphocyte migration from the lymph system into the blood system(*n)(*p).
Already disclosed practical methods of the extracorporeal processings of AIDS are reviewed following the classified items shown in our Japanese patent application, Hei4-285835.
Electromagnetic wave.
Whole blood/fraction; UV, Sun spectrum light(*m)
Leukocyte fraction; UV, Laser(*n)
Radiation.
Leukocyte fraction; X-ray(*n)
Chemicals, Pharmaceuticals
Strong inactivation agent(*k) (*m) (No actual agent is presented.)
Formaldehyde(*m)
Diethyl ether(*q)
Oxidized/peroxidized lipoprotein(*r)
ClO2, Cl2O3(*h)
Photochemical reaction with psoralen(*d)
Adsorption, adhesion, phase transition
Whole blood: Immunoadsorption column(*l)(*o)
Whole/fraction: Infectious adhesion column (No prior art)
Plasma or serum: Compounding virus with water-soluble high polymer substances having cation base such as polyvinyl pyridinum and transferring to solid phase.(*j)
Heating
Whole blood; 41xcx9c42.5xc2x0 C., 1xcx9c4 hrs.(*n)
Whole blood; 38xcx9c45xc2x0 C.(*f)
Whole blood; 56xc2x0 C., 30 min.(*g)
Plasma; 40xcx9c70xc2x0 C.(*e)
Leukocyte fraction; 39xc2x0 C. for immunostimulation(*m)
Red cell fraction; 43xc2x0 C. for killing virus(*m)
Whole blood; 41xcx9c42.5xc2x0 C., pH=7.1xcx9c7.3(*p)
Pressure
Whole blood; 2000 atm, 2xc2x0 C.xcx9c5000 atm, 25xc2x0 C.(*i)
Acoustic vibration, mechanical shear
Whole blood; mechanical shear(*n)
Acoustic vibration (no prior art)
Electric current
Whole blood or fraction: generating Ag ion from Ag electrode and sterilizing pathogenics with the ion (*s).
Electromagnetic field (no prior art)
Such anticoagulants as heparin, dextran, natrium nitrate, natrium citrate, etc., and/or such volume extender (thinner) as saline are infused after blood is withdrawn. Neutralizing the anticoagulant and/or removing excess saline by dialysis or diuresis are disclosed.
Removing or neutralizing excess agents and toxic substances generated during the process and conditioning temperature, pH, electrolytes, etc., are also disclosed as the post processing.
*a Shinichi Oka, Tokyo Medical Science Institute, NHK News, August 1993.
*b Nobuyuki Kobayashi, xe2x80x9cInfection of Virus, AIDS, and Cell Death,xe2x80x9d Nikkei Science June 1993: 34-41 and xe2x80x9cHIV Infection and Cell Death,xe2x80x9d Experimental Medicine Vol. 11, No. 5 (1993): 121-29.
*c Jean Claude Amelsen, xe2x80x9cProgrammed Cell Death and AIDS,xe2x80x9d Immunology Today 13 (1992): 388-91.
*d Emile Bisaccia et al., xe2x80x9cExtracorporeal Photopheresis in the Treatment of AIDS Related Complex,xe2x80x9d Annals of Internal Medicine 113 (1990): 270-75.
*e Japan Laid-open Pat., Sho63-240873 3/1987, Joh et al.
*f Japan Laid-open Pat., Sho63-240874 3/1987, Joh et al.
*g Japan Laid-open Pat., Sho63-252162 4/1987, Ura.
*h Japan Laid-open Pat., Hei2-274253 2/1990, Camen et al.
*i Japan Laid-open Pat., Hei3-188872 12/1989, Iwakura et al.
*j Japan Laid-open Pat., Hei4-342536 3/1991, Onishi et al.
*k U.S. Pat. No., 4,381,004 January/1981, Babb.
*l U.S. Pat. No., 4,824,432 March/1986, Skurkovich et al.
*m U.S. Pat. No., 4,908,014 September/1988, Kroyer.
*n U.S. Pat. No., 4,950,225 September/1988, Davidner et al.
*o U.S. Pat. No., 5,037,649 January/1989, Balint, Jr., et al.
*p U.S. Pat. No., 5,104,373 July/1990, Davidner et al.
*q U.S. Pat. No., 5,116,307 July/1990, Collins.
*r U.S. Pat. No., 5,192,264 October/1990, Fossel.
*s U.K. Pat. Appli. 2 189 677A April/1986, Swift
5. Problems of Prior Art
Chemical treatment has not yet succeeded. Such reverse transcription inhibitors as AZT, ddI, ddC can only delay death.
Extracorporeal blood processing has been introduced as a compelling need but has not yet been successful either. Blood processing can remove, inactivate or kill the HIVs and infected cells in blood. However, infected cells especially stay or stick to some tissue in lymphonodes, brain, etc., and their migration into the circulatory system will not be so great as expected with the administration of heparin. Most of the infected cells will stay outside of the blood stream and cannot be effectively processed with the extracorporeal blood processing only.
1. The First Method
The first method includes:
administering such Trigger Factor as TNF, anti-Fas antibody, which stimulate the proliferation control region of provirusxe2x80x94DNA in the infected cellxe2x80x94to increase proliferation rate, shorten the cell life (up to the programmed death or apoptosis) and causing the transition from latent period to ARC-AIDS period and also administering such new-infection suppressors or reverse transcription inhibitors as AZT, ddI or ddC against the proliferated free HIVs during the apoptosis process and administering as required certain agent such as heparin to induce migration of the infected cells from lymphatic system to circulatory system; and
continuing extracorporeal blood processing to kill or remove free HIVs in blood until all the infected cells die away.
The necessary processing time will be 10xcx9c24 hours, as the DNA coagulation of the infected cell is recognized in three hours after contact with TNF. Therefore, larger dose of TNF, AZT, etc., in such a short amount of time will be allowed.
It is sufficient to process only free HIVs in blood, but it is also preferable to kill or remove infected cells in blood. This processing of infected cells can shorten the necessary processing time and reduce possibility of new infection.
This treatment is preferably applied to a patient in the latent period but can be applied to a patient in ARC and AIDS period, where the dosage of Trigger Factor may be reduced or omitted, as the Trigger Factor has already been developed in the patient by AIDS.
New methods of extracorporeal blood processing by the inventors are presented as shown below; however, the extracorporeal blood processing can be a prior art.
Pharmaceuticals effective in vitro selectively to infected cells but not yet allowed in vivo administration are listed.
Trigger Factor can be proposed as well to use for the extracorporeal processing of infected cells if the pharmaceuticals, cited above are not effective. Surviving liquid, saline plus cell-cultivating agents, is used as carrier liquid. In this case, the proliferated free HIVs existing in the liquid are easily separated by filtration and rendered to such killing process as heating. The liquid can be used recirculating after the killing process.
Infectious adhesion between gp120 of HIVs/infected cells and CD4 is utilized to remove HIVs/infected cells instead of immunoadsorption, which is not effective to HIVs because of very rapid mutation and resulted variety of the stocks. Complexed with a certain high polymer or adhered to CD4-coated electroconductive or magnetic-particles, HIVs and infected cells are modified and removed by transition from the liquid phase to the solid phase through filtration, centrifugation or electromagnetic force.
Heating to higher temperature in a very short time similar to milk pasteurization with microwave device, for example, is disclosed.
Acoustic vibration, yielding mechanical shear, cavitation and heating with an ultrasound device are also presented for killing and inactivating HIVs and infected cells.
Periodically, polarity-changed DC or AC current is used to generate Cl2 or the others from NaCl, KCl in blood, and the Cl2 or the others are used to kill HIVs and infected cells.
Electromagnetic wave is used to resonate the intramolecular oscillation of HIVs or infected cells and destroy them.
The wave is also used to remove or heat the CD4-coated electroconductive or magnetic particles on the surface of which HIVs and infected cells are adhered.
2. The Second Method
The second method includes:
administering the electroconductive and/or magnetic micro particles, coated with such infectiously adhesive substance as CD4 to HIVs and infected cells; and
killing the HIVs and infected cells adhered to the micro particles by heating the micro particles with electromagnetic wave applied externally to a patient.